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Comprehensive qPCR profiling of gene expression in single neuronal cells

Citri, A., Pang, Z.P., Sudhof, T.C., Wernig, M., Malenka, R.C.

A major challenge in neuronal stem cell biology lies in characterization of lineage-specific reprogrammed human neuronal cells, a process that necessitates the use of an assay sensitive to the single-cell level. Single-cell gene profiling can provide definitive evidence regarding the conversion of one cell type into another at a high level of resolution. The protocol we describe employs Fluidigm Biomark dynamic arrays for high-throughput expression profiling from single neuronal cells, assaying up to 96 independent samples with up to 96 qPCR probes (equivalent to 9216 reactions) in a single experiment, which can be completed within 2–3 days. The protocol enables simple and cost-effective profiling of several hundred transcripts from a single cell, and could have numerous utilities.

Citation

Citri, A., Pang, Z.P., Sudhof, T.C., Wernig, M., Malenka, R.C. "Comprehensive qPCR profiling of gene expression in single neuronal cells" Nature Protocols (2011): 118-27